Substituted pyrazolopyrimidines as glucocerebrosidase activators

ABSTRACT

in which variables R1-R7 and X are described in the application.

CROSS REFERENCE TO RELATED APPLICATION

This application claims priority from U.S. provisional patentapplication No. 61/420,946, filed Dec. 8, 2010, which is herebyincorporated by reference in its entirety.

STATEMENT OF GOVERNMENT SUPPORT

This invention was made in part with government support from theNational Institutes of Health. The government has certain rights in thisinvention.

BACKGROUND

Gaucher disease is a rare disease affecting 1 in 40,000 babies born witha particular high frequency in the Ashkenazi Jews of eastern Europeandescent (about 1 in 800 live births). It is caused by inherited geneticmutations in the GBA (glucosidase, beta acid) gene, which result inreduced activity of glucocerebrosidease (GCase or acidbeta-glucocerebrosidase), an enzyme present in cellular organellescalled lysosomes, responsible for the breakdown of a fatty materialcalled glucocerebroside (or glucosyl ceramide). The accumulation of thislipid inside cells causes them to swell abnormally creating problemsthroughout the body. The disease has been categorized into three types:Neuronopathic (types 2, 3) and non-neuronopathic (type 1) with mild tosevere symptoms that can appear at anytime from infancy to adulthood.Clinical manifestations include enlarged spleen/liver, anemia, lack ofplatelets, neurodegeneration, and bone disease with varying severitydepending on the type of disease and time of diagnosis. The reduction inGCase activity has been attributed to the lack of protein in thelysosome. After production in the endoplasmic reticulum (ER) proteinsthat do not fold properly are degraded in the ER and not transported tothe lysosome where they can hydrolyze glucocerebroside.

Existing treatment options for Gaucher disease include enzymereplacement (CEREZYME) or substrate reduction therapy (ZAVESCA) whichcost between $100,000 to >$200,000 per year. The development of theiminosugar isofagomine (PLICERA) as a molecular chaperone was haltedafter Phase 2 clinical trials showed an increase in the amount of GCasein white blood cells but a lack in the reduction of visceral symptoms.Thus there is an unmet need for the development of novel chaperonetherapy for Gaucher disease. The present disclosure fulfills this needand provides additional advantages set forth in the followingdisclosure.

SUMMARY

Described herein are substituted pyrazolopyrimidines anddihydropyrazolopyrimidines and related compounds, their methods ofmanufacture, compositions containing the described compounds, andmethods of use of the described compounds. Thus in a first aspect, acompound of Formula (I) and the pharmaceutically acceptable salts of acompound of Formula (I) is provided

wherein the ring

is a ring system of the formula(i)

in which R₅ is an optionally substituted vinyl group and R₆ and R₇ carrythe definitions set forth below, or(ii)

in which R₅, R₆, and R₇ carry the definitions set forth below.

R₁ is C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, (mono- or bicycliccarbocycle)C₀-C₄alkyl or (mono- or bicyclic heterocycle)C₀-C₄alkyl, eachof which R₁ is unsubstituted or substituted with one or moresubstituents independently chosen from halogen, hydroxyl, cyano, nitro,amino, —CHO, —COOH, C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆alkoxy,C₂-C₆alkanoyl, (mono- or di-C₁-C₆alkylamino)C₀-C₄alkyl, mono- ordi-C₁-C₆alkylcarboxamide, C₁-C₆alkylester, C₁-C₆alkylthio,C₁-C₆alkylsulfonyl, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, and with 0 or 1substituents chosen from Y—Z— where Z is a covalent bond, C₁-C₄alkylene,—S—, —O—, —NR—, —C(O)—, —NHC(O)—, or —C(O)NH—, where R is hydrogen orC₁-C₄alkyl, and Y is phenyl or pyridyl, each of which is unsubstitutedor substituted with 1 to 3 substituents independently chosen fromhalogen, hydroxyl, cyano, nitro, amino, C₁-C₄alkyl, C₁-C₄alkoxy,trifluoromethyl, difluoromethyl, and trifluoromethoxy; and R₂ ishydrogen, C₁-C₆alkyl, C₃-C₇cycloalkyl, and (phenyl)C₀-C₂alkyl.

Or, R₁ and R₂ are joined to form a 5- to 7-membered heterocycloalkylring having 0 or 1 additional heteroatoms chosen from N, O, and S, which5- to 7-membered heterocycloalkyl ring is optionally fused to a phenylor pyridyl; which 5- to 7-membered heterocycloalkyl ring isunsubstituted or substituted with one or more substituents independentlychosen from halogen, hydroxyl, C₁-C₂alkyl, and C₁-C₂alkoxy.

R₃ is hydrogen or C₁-C₂alkyl.

R₅ is halogen, hydroxyl, amino, cyano, C₁-C₄alkyl, vinyl, cyclopropyl,cyclopropylidenyl, C₁-C₄alkoxy, difluoromethyl, trifluoromethyl, orphenyl.

R₆ is halogen, hydroxyl, C₁-C₄alkyl, or C₁-C₄alkoxy.

R₇ is halogen, hydroxyl, amino, cyano, C₁-C₄alkyl, C₁-C₄alkoxy,difluoromethyl, or trifluoromethyl, or

R₇ is phenyl or a 5- to 7-membered heterocycloalkyl ring having 1 or 2heteroatoms chosen from N, O, and S, each of which R₇ is directlyattached via a covalent bond or attached via a C₁-C₄ alkyl, C₁-C₄alkoxy,or C₁-C₄alkylamino group, and each of which R₇ is unsubstituted orsubstituted with 1 to 3 substituents independently chosen from halogen,hydroxyl, C₁-C₄alkyl, C₁-C₄alkxoy, (mono- anddi-C₁-C₂alkylamino)C₀-C₄alkyl, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy; or

R₆ and R₇ are taken together to form a 5- or 6-membered carbocyclic ringwith no additional points of unsaturation, which ring is unsubstitutedor substituted with 1 to 3 substituents independently chosen fromC₁-C₂alkyl and C₁-C₂alkoxy.

In certain embodiments R₁ is not unsubstituted phenyl, dihydroindenyl,benzy[b][1,4]dioxolyl, benzo[d][1,3]dioxol-5-yl, cyclohexyl, pyridyl, orphenyl substituted with 1 or 2 substituents independently chosen fromchloro, fluoro, C₁-C₄alkyl, C₁-C₂alkoxy, acetyl, and trifluoromethyl,when R₆ is hydrogen, R₅ and R₇ are both methyl, or when R₆ is hydrogenand one R₅ and R₇ is methyl and the other is phenyl. Also R₁ is not1-(4-fluorobenzyl)-1H-pyrazol-4-yl when R₆ is hydrogen and one R₅ and R₇is methyl and the other is phenyl.

In another aspect a pharmaceutical composition comprising a compound ofFormula I, or salt thereof, together with a pharmaceutically acceptablecarrier, is provided.

Also provided herein is a method of treating Gaucher disease in apatient or preventing or reducing the severity of the symptoms ofGaucher disease in a patient having a GBA gene mutation comprisingproviding an effective amount of a compound of Formula (I) orpharmaceutically acceptable salt thereof to the patient.

Yet another aspect provides a method of increasing the amount of betaglucocerebrosidase in the white blood cells of patient having a GBA genemutation, comprising providing an effective amount of a compound ofFormula (I) to the patient.

DETAILED DESCRIPTION

Described herein are substituted pyrazolopyrimidines anddihydropyrazolopyrimidines and related compounds useful as chaperones ofglucocerebrosidase. Certain substituted pyrazolopyrimidines anddihydropyrazolopyrimidines and related compounds disclosed in thisapplication are potent and selective activators of glucocerebrosidase.These compounds are useful in treating Gaucher disease and preventingthe symptoms of Gaucher disease in persons who have a mutated GBA gene.

Terminology

Compounds are described using standard nomenclature. Unless definedotherwise, all technical and scientific terms used herein have the samemeaning as is commonly understood by one of skill in the art to whichthis invention belongs.

The terms “a” and “an” do not denote a limitation of quantity, butrather denote the presence of at least one of the referenced items. Theterm “or” means “and/or”. The terms “comprising”, “having”, “including”,and “containing” are to be construed as open-ended terms (i.e., meaning“including, but not limited to”). Recitation of ranges of values aremerely intended to serve as a shorthand method of referring individuallyto each separate value falling within the range, unless otherwiseindicated herein, and each separate value is incorporated into thespecification as if it were individually recited herein. The endpointsof all ranges are included within the range and independentlycombinable. All methods described herein can be performed in a suitableorder unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”), is intended merely to better illustrate theinvention and does not pose a limitation on the scope of the inventionunless otherwise claimed. No language in the specification should beconstrued as indicating any non-claimed element as essential to thepractice of the invention as used herein. Unless defined otherwise,technical and scientific terms used herein have the same meaning as iscommonly understood by one of skill in the art to which this inventionbelongs.

All compounds are understood to include all possible isotopes of atomsoccurring in the compounds. Isotopes include those atoms having the sameatomic number but different mass numbers. By way of general example, andwithout limitation, isotopes of hydrogen include tritium and deuteriumand isotopes of carbon include ¹⁴C, ¹³C, and ¹⁴C.

The term “substituted” means that any one or more hydrogens on thedesignated atom or group is replaced with a selection from the indicatedgroup, provided that the designated atom's normal valence is notexceeded. When the substituent is oxo (i.e., ═O), then 2 hydrogens onthe atom are replaced. When aromatic moieties are substituted by an oxogroup, the aromatic ring is replaced by the corresponding partiallyunsaturated ring. For example a pyridyl group substituted by oxo is apyridone. Combinations of substituents and/or variables are permissibleonly if such combinations result in stable compounds or useful syntheticintermediates. A stable compound or stable structure is meant to imply acompound that is sufficiently robust to survive isolation from areaction mixture, and subsequent formulation into an effectivetherapeutic agent.

Suitable groups that may be present on an “optionally substituted”position include, but are not limited to, e.g., halogen, cyano,hydroxyl, amino, nitro, oxo, azido, alkanoyl (such as a C₂-C₆ alkanoylgroup such as acyl or the like); carboxamido; alkylcarboxamide; alkylgroups, alkoxy groups, alkylthio groups including those having one ormore thioether linkages, alkylsulfinyl groups including those having oneor more sulfinyl linkages, alkylsulfonyl groups including those havingone or more sulfonyl linkages, mono- and di-aminoalkyl groups includinggroups having one or more N atoms, all of the foregoing optional alkylsubstituents may have one or more methylene groups replaced by an oxygenor —NH—, and have from about 1 to about 8, from about 1 to about 6, orfrom 1 to about 4 carbon atoms, cycloalkyl; phenyl; phenylalkyl withbenzyl being an exemplary phenylalkyl group, phenylalkoxy with benzyloxybeing an exemplary phenylalkoxy group.

A dash (“-”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent.

“Alkyl” includes both branched and straight chain saturated aliphatichydrocarbon groups, having the specified number of carbon atoms. Theterm C₁-C₂alkyl means an alkyl group having from 1 to about 2 carbonatoms, e.g., methyl and ethyl, respectively Likewise “alkenyl” is abranched or straight chain unsaturated hydrocarbon group having thespecified number of carbon atoms and at least one carbon-carbon doublebond and alkynyl is a branched or straight chain unsaturated hydrocarbongroup having the specified number of carbon atoms and at least onecarbon-carbon triple bond.

“Alkylene” is a straight or branched saturated bivalent carbon chainhaving the indicated number of carbon atoms.

“Alkylester” is an alkyl group as defined above attached through anester linkage. The ester linkage may be in either orientation, e.g., agroup of the formula —O(C═O)alkyl or a group of the formula—(C═O)Oalkyl.

“Alkylester” is an alkyl group as defined above attached through anester linkage. The ester linkage may be in either orientation, e.g., agroup of the formula —O(C═O)alkyl or a group of the formula—(C═O)Oalkyl.

“Alkylester” is an alkyl group as defined above attached through anester linkage. The ester linkage may be in either orientation, e.g., agroup of the formula —O(C═O)alkyl or a group of the formula—(C═O)Oalkyl.

“Alkanoyl” is an alkyl group as defined above, attached through a keto(—(C═O)—) bridge. Alkanoyl groups have the indicated number of carbonatoms, with the carbon of the keto group being included in the numberedcarbon atoms. For example a C₂alkanoyl group is an acetyl group havingthe formula CH₃(C═O)—.

“Alkylsulfonyl” is a group of the formula alkyl-(SO₂)—, where the alkylgroup is an alkyl group as defined above having the defined number ofcarbon atoms. An exemplary alkylsulfonyl group is methylsulfonyl.

“Alkylthio” indicates an alkyl group as defined above attached through asulfur linkage, i.e. a group of the formula alkyl-S—. Examples includeethylthio and pentylthio.

“Alkoxy” means an alkyl group, as defined above, with the indicatednumber of carbon atoms attached via an oxygen bridge.

“Cycloalkyl” is a saturated hydrocarbon ring group, having the specifiednumber of carbon atoms, usually from 3 to about 7 carbon atoms. Examplesof cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl as well as bridged or caged saturated ring groups such asnorborane or adamantane.

A “mono- or bicyclic carbocycle” is a 3- to 8-membered saturated,partially unsaturated, or aromatic ring containing only carbon ringatoms or a 6 to 11 membered saturated, partially unsaturated, oraromatic bicyclic carbocyclic ring system containing only carbon ringatoms. Unless otherwise indicated, the carbocyclic group may be attachedto its pendant group at any carbon atom that results in a stablestructure. When indicated the carbocyclic rings described herein may besubstituted on any available ring carbon if the resulting compound isstable. Carbocyclic groups include cycloalkyl groups, such ascyclopropyl and cyclohexyl; cycloalkenyl groups, such as cyclohexenyl,bridged cycloalkyl groups; and aryl groups, such as phenyl.

“Halo” or “halogen” means fluoro, chloro, bromo, or iodo.

“Heterocycloalkyl” is a saturated cyclic group having the indicatednumber of ring atoms containing from 1 to about 3 heteroatoms chosenfrom N, O, and S, with remaining ring atoms being carbon. Examples ofheterocycloalkyl groups include tetrahydrofuranyl and pyrrolidinylgroups.

“Mono- or bicyclic-heterocycle” is a 5- to 8-membered saturated,partially unsaturated, or aromatic ring containing from 1 to about 4heteroatoms chosen from N, O, and S, with remaining ring atoms beingcarbon, or a 7 to 11 membered bicyclic saturated, partially unsaturated,or aromatic heterocylic ring system, each containing at least 1heteroatom in the multiple ring system chosen from N, O, and S andcontaining up to about 4 heteroatoms independently chosen from N, O, andS in each ring of the multiple ring system. Unless otherwise indicated,the heterocyclic ring may be attached to the group it substitutes at anyheteroatom or carbon atom that results in a stable structure. Whenindicated the heterocyclic rings described herein may be substituted oncarbon or on a nitrogen atom if the resulting compound is stable. Anitrogen atom in the heterocycle may optionally be quaternized. It ispreferred that the total number of heteroatoms in a heterocyclic groupsis not more than 4 and that the total number of S and O atoms in aheterocyclic group is not more than 2, more preferably not more than 1.Examples of heterocyclic groups include, pyridyl, indolyl, pyrimidinyl,pyridizinyl, pyrazinyl, imidazolyl, oxazolyl, furanyl, thiophenyl,thiazolyl, triazolyl, tetrazolyl, isoxazolyl, quinolinyl, pyrrolyl,pyrazolyl, benz[b]thiophenyl, isoquinolinyl, quinazolinyl, quinoxalinyl,thienyl, isoindolyl, dihydroisoindolyl, 5,6,7,8-tetrahydroisoquinoline,pyridinyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl,pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, and pyrrolidinyl.

“Mono- and/or di-alkylamino” means secondary or tertiary alkyl aminogroups, wherein the alkyl groups are as defined above and have theindicated number of carbon atoms. The point of attachment of thealkylamino group is on the nitrogen. The alkyl groups are independentlychosen. Examples of mono- and di-alkylamino groups include ethylamino,dimethylamino, and methyl-propyl-amino.

“Mono- or di-alkylcarboxamide” is a group of the formula—(C═O)Nalkyl₁alkyl₂, where the alkyl₁ and alkyl₂ groups areindependently chosen alkyl groups as defined herein, attached through acarboxamide linkage. The carboxamide linkage may be in eitherorientation, e.g., —NH(C═O)— or —(C═O)NH—.

“Haloalkyl” means both branched and straight-chain alkyl groups havingthe specified number of carbon atoms, substituted with 1 or more halogenatoms, generally up to the maximum allowable number of halogen atoms.Examples of haloalkyl include, but are not limited to, trifluoromethyl,difluoromethyl, 2-fluoroethyl, and penta-fluoroethyl.

“Haloalkoxy” indicates a haloalkyl group as defined above attachedthrough an oxygen bridge (oxygen of an alcohol radical).

“Pharmaceutical compositions” means compositions comprising at least oneactive agent, such as a compound or salt of Formula I, and at least oneother substance, such as a carrier. Pharmaceutical compositions meet theU.S. FDA's GMP (good manufacturing practice) standards for human ornon-human drugs.

“Carrier” means a diluent, excipient, or vehicle with which an activecompound is administered. A “pharmaceutically acceptable carrier” meansa substance, e.g., excipient, diluent, or vehicle, that is useful inpreparing a pharmaceutical composition that is generally safe, non-toxicand neither biologically nor otherwise undesirable, and includes acarrier that is acceptable for veterinary use as well as humanpharmaceutical use. A “pharmaceutically acceptable carrier” includesboth one and more than one such carrier.

A “patient” means a human or non-human animal in need of medicaltreatment. Medical treatment can include treatment of an existingcondition, such as a disease or disorder, prophylactic or preventativetreatment, or diagnostic treatment. In some embodiments the patient is ahuman patient.

“Providing” means giving, administering, selling, distributing,transferring (for profit or not), manufacturing, compounding, ordispensing.

“Treatment” or “treating” means providing an active compound to apatient in an amount sufficient to measurably reduce any symptom of abeta-glucocerebrosidase mediated disorder, e.g., cause regression of thedisorder, liver function, reduce anemia, increase platelet count, ordecrease the rate of neurodegeneration or bone degeneration. In certainembodiments treatment of Gaucher disease may be commenced before thepatient presents symptoms of the disease.

A “therapeutically effective amount” of a pharmaceutical compositionmeans an amount effective, when administered to a patient, to provide atherapeutic benefit such as an amelioration of symptoms, e.g., an amounteffective to decrease the symptoms of Gaucher disease.

A significant change is any detectable change that is statisticallysignificant in a standard parametric test of statistical significancesuch as Student's T-test, where p<0.05.

Chemical Description

Compounds of formula I may contain one or more asymmetric elements suchas stereogenic centers, stereogenic axes and the like, e.g., asymmetriccarbon atoms, so that the compounds can exist in differentstereoisomeric forms. These compounds can be, for example, racemates oroptically active forms. For compounds with two or more asymmetricelements, these compounds can additionally be mixtures of diastereomers.For compounds having asymmetric centers, all optical isomers in pureform and mixtures thereof are encompassed. In these situations, thesingle enantiomers, i.e., optically active forms can be obtained byasymmetric synthesis, synthesis from optically pure precursors, or byresolution of the racemates. Resolution of the racemates can also beaccomplished, for example, by conventional methods such ascrystallization in the presence of a resolving agent, or chromatography,using, for example a chiral HPLC column. All forms are contemplatedherein regardless of the methods used to obtain them.

All forms (for example solvates, optical isomers, enantiomeric forms,polymorphs, free compound and salts) of an active agent may be employedeither alone or in combination.

The term “chiral” refers to molecules, which have the property ofnon-superimposability of the mirror image partner.

“Stereoisomers” are compounds, which have identical chemicalconstitution, but differ with regard to the arrangement of the atoms orgroups in space.

A “diastereomer” is a stereoisomer with two or more centers of chiralityand whose molecules are not mirror images of one another. Diastereomershave different physical properties, e.g., melting points, boilingpoints, spectral properties, and reactivities. Mixtures of diastereomersmay separate under high resolution analytical procedures such aselectrophoresis, crystallization in the presence of a resolving agent,or chromatography, using, for example a chiral HPLC column.

“Enantiomers” refer to two stereoisomers of a compound, which arenon-superimposable mirror images of one another. A 50:50 mixture ofenantiomers is referred to as a racemic mixture or a racemate, which mayoccur where there has been no stereoselection or stereospecificity in achemical reaction or process.

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,Stereochemistry of Organic Compounds (1994) John Wiley & Sons, Inc., NewYork. Many organic compounds exist in optically active forms, i.e., theyhave the ability to rotate the plane of plane-polarized light. Indescribing an optically active compound, the prefixes D and L or R and Sare used to denote the absolute configuration of the molecule about itschiral center(s). The prefixes d and 1 or (+) and (−) are employed todesignate the sign of rotation of plane-polarized light by the compound,with (−) or 1 meaning that the compound is levorotatory. A compoundprefixed with (+) or d is dextrorotatory.

A “racemic mixture” or “racemate” is an equimolar (or 50:50) mixture oftwo enantiomeric species, devoid of optical activity. A racemic mixturemay occur where there has been no stereoselection or stereospecificityin a chemical reaction or process.

“Pharmaceutically acceptable salts” include derivatives of the disclosedcompounds in which the parent compound is modified by making inorganicand organic, non-toxic, acid or base addition salts thereof. The saltsof the present compounds can be synthesized from a parent compound thatcontains a basic or acidic moiety by conventional chemical methods.Generally, such salts can be prepared by reacting free acid forms ofthese compounds with a stoichiometric amount of the appropriate base(such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, or thelike), or by reacting free base forms of these compounds with astoichiometric amount of the appropriate acid. Such reactions aretypically carried out in water or in an organic solvent, or in a mixtureof the two. Generally, non-aqueous media such as ether, ethyl acetate,ethanol, isopropanol, or acetonitrile are used, where practicable. Saltsof the present compounds further include solvates of the compounds andof the compound salts.

Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include theconventional non-toxic salts and the quaternary ammonium salts of theparent compound formed, for example, from non-toxic inorganic or organicacids. For example, conventional non-toxic acid salts include thosederived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, nitric and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic,mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic,HOOC—(CH₂)_(n)—COOH where n is 0-4, and the like. Lists of additionalsuitable salts may be found, e.g., in Remington's PharmaceuticalSciences, 17th ed., Mack Publishing Company, Easton, Pa., p. 1418(1985).

Small molecules which activate the GCase enzyme are disclosed herein.The data suggests these small molecules may be acting as chaperoneswhich help the misfolded enzyme to fold properly and be trafficked fromthe endoplasmic reticulum to the lysosome. Most small moleculechaperones described in literature are inhibitors of GCase and thus canpotentially inhibit enzyme activity in the lysosome. The presentchemical series is advantageous as the compounds do not inhibit, butrather activate GCase. The chemical class of pyrazolo[1, 5-a]pyrimidine-3carboxamides is also structurally distinct from iminosugars,often described as chaperones in literature, and holds promise towardsselectivity against other glycosidases.

In addition to compounds of Formula (I) shown above in the SUMMARYsection, Compounds of II and III, which are subformulae of Formula I,and compounds in which the variables, e.g., R₁-R₇ carry the followingdefinitions are also disclosed.

In Formula III, R_(5a) is hydrogen, C₁-C₄alkyl, C₃-C₇cycloalkyl, or 4-to 7-membered carbon attached heterocycloalkyl, having 1 or 2heteroatoms independently chosen from N, S, and O.

In certain embodiments of Formula III, R_(5a) is hydrogen orcyclopropyl.

Included herein are compounds and salts for Formula I and II in which:

R₂ is hydrogen or methyl; and

R₅ is C₁-C₄alkyl, difluoromethyl, or phenyl;

R₇ is C₁-C₄alkyl, difluoromethyl, or phenyl; and

R₅ and R₇ are not both phenyl.

Further included herein are compounds and salts for Formula I and II inwhich: R₅ and R₇ are both methyl; or

one of R₅ and R₇ is methyl and the other is phenyl; or

one of R₅ and R₇ is methyl and the other is difluoromethyl.

Included herein are compounds and salts of Formula I, II, and III inwhich: R₁ is (phenyl)C₀-C₄alkyl, (pyridyl)C₀-C₄alkyl, (pyrimidinyl)C₀-C₄alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, (pyrazolyl)C₀-C₂alkyl,(pyrrolyl)C₀-C₂alkyl, (imidazolyl)C₀-C₂alkyl, (thienyl)C₀-C₂alkyl,(furanyl)C₀-C₂alkyl, (oxazolyl)C₀-C₂alkyl, (thiazolyl)C₀-C₂alkyl,pyrolidinyl, naphthyl, quinolinyl, isoquinolinyl, tetrahydronaphthyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, tetrahydrofuranyl,piperazinyl, morpholinyl, piperidinyl, thiomorpholinyl, dihydroindenyl,benzo[b][1,4]dioxinyl, or benzo[d][1,3]dioxolyl, each of which isunsubstituted or substituted with one or more substituents independentlychosen from halogen, hydroxyl, cyano, nitro, amino, —CHO, —COOH,C₁-C₆alkyl, C₁-C₆alkoxy, C₂-C₆alkanoyl, mono- or di-C₁-C₆alkylamino,mono- or di-C₁-C₆alkylcarboxamide, C₁-C₆alkylester, C₁-C₆alkylthio,C₁-C₆alkylsulfonyl, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, and with 0 or 1substituents chosen from Y—Z— where Z is a covalent bond, C₁-C₄alkylene,—S—, —O—, —NR—, —C(O)—, —NHC(O)—, or —C(O)NH—, where R is hydrogen orC₁-C₄alkyl, and Y is phenyl or pyridyl, each of which is unsubstitutedor substituted with 1 to 3 substituents independently chosen fromhalogen, hydroxyl, cyano, nitro, amino, C₁-C₄alkyl, and C₁-C₄alkoxy.

Also included herein are compounds and salts of Formula I, II, and IIIin which: R₁ and R₂ are joined to form a 5- to 7-memberedheterocycloalkyl ring having 0 or additional heteroatoms chosen from N,O, and S, which 5- to 7-membered heterocycloalkyl ring is optionallyfused to a phenyl or pyridyl; which 5- to 7-membered heterocycloalkylring is unsubstituted or substituted with one or more substituentsindependently chosen from halogen, hydroxyl, C₁-C₂alkyl, andC₁-C₂alkoxy.

Further included herein are compounds and salts of Formula I, II, andIII in which R₁ is (phenyl)C₀-C₂alkyl, substituted with at least onesubstituent chosen from cyano, trifluoromethyl, CH₃C(O)NH—, or nR₁ iscyclohexyl, substituted with at least one trifluoromethyl, C₃-C₆alkyl;or R₁ is dihydroindenyl, quinolinyl, or isoquinolinyl; each of which R₁may be substituted with one or more substituents independently chosenfrom halogen, hydroxyl, cyano, nitro, amino, —CHO, —COOH, C₁-C₄alkyl,C₁-C₄alkoxy, C₂-C₄alkanoyl, mono- or di-C₁-C₄alkylamino, C₁-C₂haloalkyl,and C₁-C₂haloalkoxy.

Included herein are compounds and salts of Formula I, II, and III inwhich: R₂ is hydrogen or methyl; and R₇ is C₁-C₄alkyl, difluoromethyl,or phenyl. In some embodiments it is preferred that R₇ isdifluoromethyl.

Included herein are compounds and salts of Formula I, II, and III inwhich: R₂ is hydrogen or methyl; and R₇ is methyl or difluoromethyl; and

R₁ is (phenyl)C₀-C₂alkyl, (pyridyl)C₀-C₂alkyl, (cyclohexyl)C₀-C₂alkyl,pyrazolyl, furanylnaphthyl, quinolinyl, isoquinolinyl,tetrahydronaphthyl, tetrahydroquinolinyl, tetrahydroisoquinolinyltetrahydrofuranyl, morpholinyl, piperidinyl, piperazinyl,thiomorpholinyl, dihydroindenyl, benzo[b][1,4]dioxinyl, orbenzo[d][1,3]dioxolyl, each of which is unsubstituted or substitutedwith one or more substituents independently chosen from halogen,hydroxyl, cyano, nitro, amino, —CHO, —COOH, C₁-C₄alkyl, C₁-C₄alkoxy,C₂-C₄alkanoyl, mono- or di-C₁-C₄alkylamino, mono- ordi-C₁-C₄alkylcarboxamide, C₁-C₄alkylester, C₁-C₂alkylsulfonyl,trifluoromethyl, trifluoromethoxy, and difluoromethyl, and with 0 or 1substituents chosen from Y—Z— where Z is a covalent bond, C₁-C₄alkylene,—S—, —O—, —NR—, —C(O)—, —NHC(O)—, or —C(O)NH—, where R is hydrogen orC₁-C₄alkyl, and Y is phenyl or pyridyl, each of which is unsubstitutedor substituted with 1 to 3 substituents independently chosen fromhalogen, hydroxyl, C₁-C₂alkyl, and C₁-C₂alkoxy.

Compounds of Formula I have the following tautomeric formulas:

Pharmaceutical Preparations

The substituted pyrazolopyrimidines and dihydropyrazolopyrimidinesdisclosed herein can be administered as the neat chemical, but arespecifically administered as a pharmaceutical composition, for example apharmaceutical formulation comprising a substituted pyrazolopyrimidinesor dihydropyrazolopyrimidine compound of Formula I or pharmaceuticallyacceptable salt thereof, together with at least one pharmaceuticallyacceptable carrier. The pharmaceutical composition can be is formulatedas an injectable fluid, an aerosol, a cream, a gel, a tablet, a pill, acapsule, a syrup, or a transdermal patch.

The substituted pyrazolopyrimidines and dihydropyrazolopyrimidines maybe administered orally, topically, parenterally, by inhalation or spray,sublingually, transdermally, via buccal administration, rectally, as anophthalmic solution, or by other means, in dosage unit formulationscontaining conventional pharmaceutically acceptable carriers. Thepharmaceutical composition may be formulated as any pharmaceuticallyuseful form, e.g., as an aerosol, a cream, a gel, a pill, a capsule, atablet, a syrup, a transdermal patch, or an ophthalmic solution. Somedosage forms, such as tablets and capsules, are subdivided into suitablysized unit doses containing appropriate quantities of the activecomponents, e.g., an effective amount to achieve the desired purpose.

Carriers include excipients and diluents and must be of sufficientlyhigh purity and sufficiently low toxicity to render them suitable foradministration to the patient being treated. The carrier can be inert orit can possess pharmaceutical benefits of its own. The amount of carrieremployed in conjunction with the compound is sufficient to provide apractical quantity of material for administration per unit dose of thecompound.

Classes of carriers include, but are not limited to binders, bufferingagents, coloring agents, diluents, disintegrants, emulsifiers,flavorings, glidants, lubricants, preservatives, stabilizers,surfactants, tableting agents, and wetting agents. Some carriers may belisted in more than one class, for example vegetable oil may be used asa lubricant in some formulations and a diluent in others. Exemplarypharmaceutically acceptable carriers include sugars, starches,celluloses, powdered tragacanth, malt, gelatin, talc, and vegetableoils. Optional active and/or inactive agents may be included in thepharmaceutical compositions, provided that such agents do notsubstantially interfere with the activity of the hydrazone and diacylhydrazine compounds used in the pharmaceutical compositions. Theoptional active is an additional active agent that is not a compound orsalt of Formula I.

The pharmaceutical compositions can be formulated for oraladministration. These compositions contain between 0.1 and 99 weight %(wt. %) of a hydrazone or a diacyl hydrazine compound and usually atleast about 5 wt. % of a hydrazone or a diacyl hydrazine compound. Someembodiments contain from about 25 wt. % to about 50 wt. % or from about5 wt. % to about 75 wt. % of the hydrazone or diacyl hydrazine compound.

Treatment Methods

The compounds of Formula I or a salt thereof, as well as pharmaceuticalcompositions comprising the compounds, are useful for treating lysosomalstorage diseases, including Gaucher disease. The compounds of Formula Ior a salt thereof, as well as pharmaceutical compositions comprising thecompounds, are also useful for preventing the occurrence of symptoms ofa lysosomal storage disorder, such as Gaucher disease, in a patienthaving GBA gene mutation. The method of treating a lysosomal storagedisease in a patient comprises providing to the patient an effectiveamount of a compound or salt of Formula I: In an embodiment the patientis a mammal, specifically a primate, more specifically a human. Aneffective amount of a pharmaceutical composition may be an amountsufficient to inhibit the progression of a disease or disorder; or causea regression of a disease or disorder.

An effective amount of a compound or pharmaceutical compositiondescribed herein will also provide a sufficient concentration of asubstituted pyrazolopyrimidines or dihydropyrazolopyrimidines compoundwhen administered to a patient. A sufficient concentration is aconcentration of the compound or salt of Formula I in the patient's bodynecessary to prevent or combat the disorder. Such an amount may beascertained experimentally, for example by assaying blood concentrationof the compound, or theoretically, by calculating bioavailability.

Methods of treatment include providing certain dosage amounts of asubstituted pyrazolopyrimidine or dihydropyrazolopyrimidine compound orsalt to a patient. Dosage levels of each compound of from about 0.1 mgto about 140 mg per kilogram of body weight per day are useful in thetreatment of the above-indicated conditions (about 0.5 mg to about 7 gper patient per day). The amount of compound that may be combined withthe carrier materials to produce a single dosage form will varydepending upon the patient treated and the particular mode ofadministration. Dosage unit forms will generally contain between fromabout 1 mg to about 500 mg of each active compound. In certainembodiments 25 mg to 500 mg, or 25 mg to 200 mg of a compound of FormulaI are provided daily to a patient. Frequency of dosage may also varydepending on the compound used and the particular disease treated.However, for treatment of most diseases and disorders, a dosage regimenof 4 times daily or less can be used and in certain embodiments a dosageregimen of 1 or 2 times daily is used.

It will be understood, however, that the specific dose level for anyparticular patient will depend upon a variety of factors including theactivity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, route ofadministration, and rate of excretion, drug combination and the severityof the particular disease undergoing therapy.

In an embodiment, the invention provides a method of treating alysosomal storage disorder in a patient identified as in need of suchtreatment, the method comprising providing to the patient an effectiveamount of a compound of Formula I. The compounds and salts of Formula Iprovided herein may be administered alone, or in combination with one ormore other active agent.

Methods of treatment provided herein are also useful for treatment ofmammals other than humans, including for veterinary applications such asto treat horses and livestock e.g. cattle, sheep, cows, goats, swine andthe like, and pets (companion animals) such as dogs and cats.

For diagnostic or research applications, a wide variety of mammals willbe suitable subjects including rodents (e.g. mice, rats, hamsters),rabbits, primates, and swine such as inbred pigs and the like.Additionally, for in vitro applications, such as in vitro diagnostic andresearch applications, body fluids (e.g., blood, plasma, serum, cellularinterstitial fluid, saliva, feces and urine) and cell and tissue samplesof the above subjects will be suitable for use.

Examples

The following Examples are offered by way of illustration and not by wayof limitation. Unless otherwise specified, all reagents and solvent areof standard commercial grade and are used without further purification.Starting materials are available from commercial suppliers, such asSigma-Aldrich (St. Louis, Mo.), or are synthesized using procedures thatare known in the art.

Example 1. Synthetic Scheme for Preparing SubstitutedPyrazolopyrimidines and Dihydropyrazolopyrimidines

The synthetic sequence (Scheme 1) used to generate the substitutedpyrazolopyrimidines and dihydropyrazolopyrimidines provided hereinstarts with the condensation of a 1,3-diketone 6 with ethyl3-amino-1H-pyrazole-4-carboxylate 7 in acetic acid. Such reactions havebeen reported previously. See for example WO 2008/134035 and Huppatz, J.L. Aust. J Chem. 1985, 38, 221-230. Initial efforts to simplify theevaluation of the amide SAR centered on the use of acetylacetone as thediketone to eliminate the formation of regioisomers (R₁═R₂═CH₃). Withunsymmetrical ketones a mixture of regioisomers (8, 9) forms which istypically separated by chromatography. Ester hydrolysis then provides anacid, which is coupled with amines or anilines to generate compounds(10, 11) for biological analysis.

Example 2. Synthesis ofN-(4-ethynylphenyl)-5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamideStep 1. Synthesis of 5,7-Dimethylpyrazolo [1,5-a]pyrimidine-3-carboxylicacid (12)

Pentane-2,4-dione (1.46 ml, 14.2 mmol), ethyl3-amino-1H-pyrazole-4-carboxylate (2.00 g, 12.9 mmol) were heated in asealed tube with acetic acid (10 ml) at 110° C. overnight. The reactionreached completion by LCMS (LC-MS: rt (min)=3.08). The acetic acid wasremoved by blowing air with the flask being heated to 75° C. The cruderesidue of ethyl 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxylate(assumed to be 12.89 mmol) was suspended in MeOH (15 ml) and treatedwith 7.2 M sodium hydroxide (5.37 ml, 38.7 mmol). The mixture was heatedto 80° C. (at this temperature the solid dissolved) and then stirred for3 h. The reaction was cooled and the neutralized to pH 6-7. The slurrywas filtered via a Buchner funnel under house vacuum, the solid residuewas washed with water and then diethyl ether to obtain5,7-dimethylpyrazolo [1,5-a]pyrimidine-3-carboxylic acid (1.4 g, 7.3mmol, 57% yield). LC-MS: rt (min)=2.61. 1H NMR (400 MHz, DMSO-d₆) δ ppm2.57 (s, 3H), 2.71 (s, 3H), 7.10 (s, 1H), 8.50 (s, 1H).

Step 2. Synthesis ofN-(4-Ethynylphenyl)-5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide(13)

5,7-Dimethylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (722 mg, 3.78mmol), 4-ethynylaniline (442 mg, 3.78 mmol), and HATU (1436 mg, 3.78mmol) were taken up in DMF (10 ml) and then treated withdiisopropylethylamine (1.979 ml, 11.33 mmol). The contents stirred atroom temperature overnight. The product had precipitated from reactionmixture. The reaction was diluted with water, filtered through a Buchnerfunnel under house vacuum. The residue was washed with water (×2), thenCH₂Cl₂, diethyl ether, and air dried to obtainN-(4-ethynylphenyl)-5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide(500 mg, 1.72 mmol, 46% yield). LC-MS: rt (min)=3.65. 1H NMR (400 MHz,DMSO-d₆) δ ppm 2.71 (s, 3H), 2.77 (s, 3H), 4.11 (s, 1H), 7.21 (s, 1H),7.49 (d, J=8.6 Hz, 2H), 7.77 (d, J=8.6 Hz, 2H), 8.65 (s, 1H), 10.31 (s,1H).

Example 3. Assay Protocol for Glucocerebrosidase Activity

qHTS Assay for Inhibitors and Activators of N370S glucocerebrosidase asa Potential Chaperone Treatment of Gaucher Disease: Primary ScreenConfirmation

This is a fluorogenic enzyme assay with4-methylumbelliferyl-beta-D-glucopyranoside as the substrate and N370Sglucocerebrosidase from human spleen homogenate as the enzymepreparation. Upon the hydrolysis of this fluorogenic substrate, theresulting product, 4-methyllumbelliferone, can be excited at 365 nm andemits at 440 nm which can be detected by a standard fluorescence platereader. Data were normalized to the controls for basal activity (withoutenzyme) and 100% activity (with enzyme). The AC50 values were determinedfrom concentration-response data modeled with the standard Hillequation.

Human spleen homogenate is prepared as follows by homogenizing spleenwith assay buffer containing 50 mM citric acid (titrated with potassiumphosphate to pH 5.0), 100 mM potassium chloride, 10 mM sodium chloride,1 mM magnesium chloride, 0.01% Tween-20. The protein concentration ofthe human spleen homogenate is approximated 1.35 ug/ul.

The assay is performed in 1536 well plates according to the followingprotocol. (1) 2 ul spleen homogenate (27 ug final) is added to eachwell. (2) 23 nL test compound in DMSO solution is added to each well.The final concentration of compound in the test well is 0.5 nM to 58 uM.(3) 2 ul of substrate (1 mM final) are added to each well. (4) Incubateassay plates at 37° C. for 40 min. (5) Add 2 ul stop solution (1M NaOHand 1M Glycine mixture, pH 10) to each well. (6) Detect the assay platein a ViewLux plate reader (PerkinElmer) using 365 nm excitation withemission at 440 nm.

Example 4. Chaperone Activity ofN-(4-ethynylphenyl)-5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide(NGC00188758-01)

Chaperone activity of NGC00188758-1 (1 μM and 5 μM), isofagomine (1 μM),and a vehicle control were evaluated in a N370S patient derivedfibroblast for chaperone activity. GCase was visualized with red stain(anti GCase R386 antibody), lysosome with green (LAMP), and nucleus withblue (DAPI). An overlay of all stains shows bright yellow color (due tooverlap of red and green color) color indicating the presence of GCasein the lysosome. All laser settings were held constant across all of thefields imaged. Both isofagomine and NGC00188758-1 showed significantlyincreased GCase activity in the lysosome relative to vehicle control.

Example 5. Selectivity Assay: qHTS Assay for Inhibitors and Activatorsof N370S Glucocerebrosidase as a Potential Chaperone Treatment ofGaucher Disease: Alpha-Glucosidase Counterscreen

To characterize compound selectivity, selected hits from the primaryscreen were screened against purified alpha-glucosidase, a related sugarhydrolase. Alpha-glucosidase is responsible for hydrolysis of terminal,non-reducing 1,4-linked alpha-D-glucose residues with release ofalpha-D-glucose. This is a fluorogenic enzyme assay using4-methylumbelliferyl-alpha-D-pyranoside as the substrate and humanalpha-glucosidase as the enzyme preparation. Upon the hydrolysis of thisfluorogenic substrate, the resulting product, 1.4-methyllumbelliferone,can be excited at 365 nm and emits at 440 nm. Emission is detected by astandard fluorescence plate reader. Data were normalized to the controlsfor basal activity (without enzyme) and 100% activity (with enzyme). TheAC50 values were determined from concentration-response data modeledwith the standard Hill equation.

The assay is performed in 1536 well plates according to the followingprotocol. (1) Add 2 ul/well of human alpha-glucosidase enzyme (4 nMfinal) in assay buffer (50 mM citric acid (titrated with potassiumphosphate to pH 5.0), 0.005% Tween-20, pH 5.0) to each well. (2) Add 23nL compounds in DMSO solution. The final compound titration is 0.7 nM to77 uM. (3) Add 1 ul of substrate (400 uM final). (4) Incubate at roomtemperature for 20 min. (5) Add 2 ul stop solution (1M NaOH and 1MGlycine mixture, pH 10) (6) Detect the assay plate in a ViewLux platereader (PerkinElmer) with Ex=365 nm and Em=440 nm.

Example 6. Lysosomal Translocation Assay: Chaperone Activity in GaucherFibroblasts after Multi-Day Incubation with Compound

This assay quantitates translocated glucocerebrosidase protein inpatient-derived fibroblasts following extended compound incubation. Thefibroblasts tested in this experiment were homozygous for N370Sglucocerebrosidase.

Primary dermal fibroblasts derived from skin biopsies from twopreviously described N370S/N370S Gaucher patients (Goker-Alpan et al,2008) and a control were seeded in Lab-Tek 4 chamber slides (FisherScientific, Pittsburgh, Pa.). After compound treatment fibroblasts werefixed in 3% paraformaldehyde. The cells were permeabelized with 0.1%Triton-X for 10 min. and blocked in PBS containing 0.1% saponin, 100 μMglycine, 0.1% BSA and 2% donkey serum followed by incubation with mousemonoclonal anti-LAMP1 or LAMP-2 (1:100, Developmental Studies Hybridomabank, University of Iowa, Iowa City, IA) and the rabbit polyclonalanti-GCase R386 antibody (1:500). The cells were washed and incubatedwith secondary donkey anti-mouse or anti-rabbit antibodies conjugated toALEXA-488 or ALEXA-555, respectively (Invitrogen, Carlsbad, Calif.),washed again, and mounted in VectaShield with DAPI (Vector Laboratories,Burlingame, Calif.).

Cells were imaged with a Zeiss 510 META confocal laser-scanningmicroscope (Carl Zeiss, Microimaging Inc., Germany) using an Argon (458,477, 488, 514 nm) 30 mW laser, a HeNe (543 nm) 1 mW laser, and a laserdiode (405 nm). Low and high magnification images were acquired using aPlan-Apochromat 20×/0.75 objective and a Plan-Apochromat 100×/1.4 oilDIC objective, respectively. Images were taken with the same lasersettings and all the images are collapsed z-stacks. Images suggestedsignificant translocation of glucocerebrosidase protein to lysosomes incompound treated fibroblasts relative to untreated fibroblasts.

Example 8. Additional Compounds

The following compounds are prepared according to the proceduresprovided in Examples 1 and 2. Those of skill in the art will recognizethat reagents and reaction conditions will need to be varied to achievethe listed compounds. Such variations will be readily apparent to thoseof skill in the art of organic chemical synthesis.

LCMS retention time data was obtained as follows:

Method 1. Column: Phenomenex Luna C18 (3 micron, 3×75 mm). Run time: 8min. Gradient: 4% to 100% Acetonitrile in water over 7 min. Mobilephase: Acetonitrile (0.025% TFA), water (0.05% TFA). Flow rate: 1mL/min. Temperature: 50° C. UV wavelength: 220 nm, 254 nm

2. Method 2: detection UV214, Gemini column, Solvent A is water, SolventB is 90% acetonitrile, 10% water; 0.1% AcOH modifier. Gradient initialcondition is 100% A with hold time 0.5 min; gradient time is 3 min.Gradient final conc. is 100% B. gradient final hold is 0.5 min. Run time5 min. Flow ate 1.5 ml/min.

Method 3. detection UV220, Phenomenex Luna 2.5 micron C18 100×2.00 mmcolumn, Solvent A is water (0.05% TFA), Solvent B is acetonitrile(0.025% TFA). Gradient initial condition is 95% A, 5% B with hold time0.1 min; gradient time is 2.1 min. Gradient final conc. is 100% B, 5% A.gradient final hold is 0.5 min. Then another hold for further 0.3 minwith 98% A, 2% B. Run time 3 min. Flow ate 0.30 to 0.5 ml/min. The LCMSretention times are from Method 1 unless indicated otherwise.

LCMS ret. time Cmpd. # Structure Name AC50 (min.) NCGC00188758- 01

N-(4-ethynylphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 0.3659 5.784 NCGC00188783- 01

N-(3,4-dimethylphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 1.4566 6.094 NCGC00188787- 01

5,7-dimethyl-N-(4- (trifluoromethyl)phenyl) pyrazolo[1,5-a]pyrimidine-3- carboxamide 1.4566 6.434 NCGC00188776- 01

N-(3-fluoro-4-methoxy- phenyl)- 5,7-dimethylpyrazolo[,15-a]pyrimidine-3-carboxamide 1.4566 5.519 NCGC00188772- 01

5,7-dimethyl-N-p- tolylpyrazolo[1,5-a] pyrimidine- 3-carboxamide 1.63435.823 NCGC00188782- 01

N-(3-chloro-4- methylphenyl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 1.8338 6.437 NCGC00188756- 01

N-(4-bromophenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide2.0575 6.208 NCGC00188780- 01

N-(3-chloro-4-methoxy- phenyl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 2.0575 5.793 NCGC00229708- 01

N-(4-tert-butylcyclohexyl)-7- methyl-5-phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 2.0575 7.439 MLS000708974-01

7-(difluoromethyl)-N-(4- ethylphenyl)-5- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide 2.2387 2.47 Method 2 NCGC00188764- 01

N-(4-tert-butylphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 2.3086 6.745 NCGC00182141- 01

N-(4-sec-butylphenyl)-7- (difluoromethyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.5119 2.67 Method 3NCGC00182160- 01

7-(difluoromethyl)-N-(2- ethoxyphenyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.5119 2.44 Method 3NCGC00182179- 01

N-(4-bromo-2-fluorophenyl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.5119 2.71 Method 3NCGC00187969- 01

N-(2,3-dihydro-1H- inden-5-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 2.5902 6.328 NCGC00182133- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(3,4- dimethylphenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 3.1623 2.59 Method 3NCGC00182171- 01

N-(5-chloropyridin-2-yl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 3.1623 2.58 Method 3NCGC00182172- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(2,5- difluorophenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 3.1623 2.57 Method 3NCGC00187970- 01

N-(4-ethylphenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide3.2609 6.231 MLS000662187-01

7-(difluoromethyl)-N-(1-(4- fluorobenzyl)-1H-pyrazol-4-yl)-5-phenylpyrazolo[1,5- a]pyrimidine-3-carboxamide 3.2609 2.34 Method2 NCGC00229713- 01

N-(3,4-dichlorophenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 3.6588 6.462 NCGC00182174- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(2,3-dihydro-1H-inden-5-yl)-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 3.9811 2.64 Method 3 NCGC00182166- 01

N-(2,4-difluorophenyl)-7- methyl-5-methylene-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 3.9811 NCGC00188755- 01

N-(4-tert-butylcyclohexyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 4.1053 6.939 MLS000662237-01

N-(4-sec-butylphenyl)-7- (difluoromethyl)-5- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide 5.0119 2.76 Method 2 NCGC00188766- 01

N-(4-fluorophenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide5.7988 5.581 NCGC00229707- 01

N-(4-tert-butylcyclohexyl)-7- methyl-5-phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 5.7988 7.298 NCGC00187967- 01

N-(3,5-difluorophenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 6.1783 6.129 NCGC00054856- 02

N-(benzo[d][1,3]dioxol-5-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 6.5064 5.329 NCGC00188771- 01

5,7-dimethyl-N-m- tolylpyrazolo[1,5-a] pyrimidine-3-carboxamide 6.50645.841 NCGC00229715- 01

N-(3,4-dichlorophenyl)-7- methyl-5-phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 6.5064 7.257 MLS000765493-01

N-(3,4-dichlorophenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 7.0795 2.66 Method 2 NCGC00182186- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(2- (phenylthio)phenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 7.0795 2.72 Method 3NCGC00229719- 01

N-(2,3-dihydro-1H- inden-5-yl)-7-methyl- 5-phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 8.1911 7.025 NCGC00187975- 01

5,7-dimethyl-N- phenylpyrazolo[1,5- a]pyrimidine-3-carboxamide 8.19115.522 NCGC00188768- 01

N-(4-methoxyphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 8.1911 5.328 NCGC00182167- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(2,3-dihydrobenzo[b][1,4]dioxin-6- yl)-4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carboxmaide 8.9125 2.37 Method 3 NCGC00229717- 01

N-(4-ethylphenyl)-5- methyl-7- phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 9.1905 6.884 NCGC00182142- 01

7-(difluoromethyl)-N-(3- ethylphenyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 10 2.50 Method 3NCGC00182146- 01

N-(2,4-dichlorobenzyl)-7- (difluoromethyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 10 2.48 Method 3NCGC00182165- 01

methyl 2-(7-methyl-5- methylene-4,5- dihydropyrazolo[1,5-a]pyrimidine-3- carboxamido)benzoate 10 2.36 Method 3 NCGC00188773- 01

N-(2,3- dihydrobenzo[b][1,4]dioxin-6- yl)-5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 10.3119 5.301 NCGC00187971- 01

N-cyclohexyl-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide10.3119 5.551 NCGC00182182- 01

N-(4-tet-butylcyclohexyl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 11.2202 2.83 Method 3NCGC00182154- 01

N-(4-(diethylamino)phenyl)-7- (difluoromethyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 11.2202 1.75 Method3 NCGC00188779- 01

N-(2-fluoro-4-methylphenyl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 11.5702 6.112 NCGC00182170- 01

N-cyclopentyl-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 12.5893 2.39 Method 3NCGC00182175- 01

N-cyclohexyl-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 12.5893 2.48 method 3NCGC00182139- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(6- methylpyridin-2-yl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 12.5893 2.15 Method 3NCGC00182185- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(3- (phenylamino)phenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 12.5893 2.55 Method 3NCGC00182188- 01

N-(2-bromophenyl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 12.5893 2.60 Method 3NCGC00182186- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(2- (phenylthio)phenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 12.982 2.72 Method 3NCGC00188778- 01

N-(2,4-dimethylphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 12.982 6.112 NCGC00229714- 01

N-(4-ethylphenyl)-7-methyl-5- phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 12.982 6.914 NCGC00182162- 01

7-(difluoromethyl)-N-(2- methoxybenzyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 14.1254 2.26 Method3 NCGC00182190- 01

5-cyclopropylidene-N-(2- (phenylthio)phenyl)-7- (trifluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 14.1254 2.76 Method 3NCGC00182176- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(1,2,3,4-tetrahydronaphthalen-1-yl)-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 14.254 NCGC00182148- 01

N-(2-chlorobenzyl)-7- (difluoromethyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 14.1254 2.35 Method3 NCGC00182132- 01

methyl 2-(5-cyclopropylidene- 7-(difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3- carboxamido)benzoate 15.8489 2.54Method 3 NCGC00182159- 01

N-(4-chlorophenethyl)-7- (difluoromethyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 15.8489 2.40 Method3 NCGC00182161- 01

7-(difluoromethyl)-5- methylene-N-(2- (trifluoromethyl)benzyl)-4,5-dihdyropyrazolo[1,5- a]pyrimidine-3-carboxamide 15.8489 2.39 Method 3NCGC00182197- 01

N-hexyl-7-methyl-5-methylene- 4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 15.8489 2.47 Method 3 NCGC00182198- 01

7-methyl-5-methylene-N-(5- methylpyridin-2-yl)-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 15.8489 1.78 Method 3 NCGC00188770- 01

5,7-dimethyl-N-o- tolylpyrazolo[1,5-a]pyrimidine- 3-carboxamide 16.34335.730 NCGC00188762- 01

5,7-dimethyl-N-(quinolin-3- yl)pyrazolo[1,5-a]pyrimidine- 3-carboxamide16.3433 4.266 NCGC00187965- 01

N-(3-chlorobenzyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide18.3375 5.639 NCGC00229712- 01

5,7-dimethyl-N-(2- (phenylthio)phenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide 18.3375 6.590 NCGC00188763- 01

5,7-dimethyl-N-(quinolin-2- yl)pyrazolo[1,5-a]pyrimidine- 3-carboxamide20.575 4.637 NCGC00188769- 01

N-(4-cyanophenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide20.575 5.441 NCGC00182147- 01

7-(difluoromethyl)-N-(2- methoxyphenethyl)-5- methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 25.1189 2.31 Method 3NCGC00188754- 01

N-(4-tert-butylcyclohexyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 25.9024 6.816 NCGC00188781- 01

N-(2,4-dimethoxyphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 25.9024 5.483 NCGC00229716- 01

N-(3,5-dichlorophenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 25.9024 6.715 NCGC00182145- 01

N-sec-butyl-7- (difluoromethyl)- 5-methylene-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 31.6228 2.22 Method 3 NCGC00182151- 01

N-(2-chlorophenethyl)-7- (difluoromethyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 31.6228 2.38 Method3 NCGC00182152- 01

(7-(difluoromethyl)-5- methylene-4,5- dihydropyrazolo[1,5-a]pyrimidin-3-yl)(4- phenylpiperazin-1- yl)methanone 31.6228 2.08 Method3 NCGC00187978- 01

N-isobutyl-5,7- dimethylpyrazolo[,15- a]pyrimidine-3-carboxamide 32.60925.075 NCGC00188785- 01

N-(3-acetylphenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide32.6092 5.266 NCGC0229711- 01

N-(4-tert-butylcyclohexyl)-5- methyl-7-phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 32.6092 7.407 NCGC00182144- 01

(7-(difluoromethyl)-5- methylene-4,5- dihydropyrazolo[1,5-a]pyrimidin-3-yl)(4- (pyridin-2- yl)piperazin-1-yl)methanone 35.48131.59 NCGC00188786- 01

5,7-dimethyl-N-(3- (trifluoromethyl) phenyl)pyrazolo[1,5-a]pyrimidine-3- carboxamide 41.0526 6.364 NCGC00187976- 01

N-benzyl-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide 51.68215.189 NCGC00188759- 01

N-(furan-2-ylmethyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 51.6821 4.723 NCGC00188774- 01

N-(4-methoxy-2- methylphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 51.6821 5.530 NCGC00188784- 01

N-(4-acetylphenyl)-5,7- dimethylpyraozlo[1,5- a]pyrimidine-3-carboxamide163.4332 5.250 NCGC0229718- 01

5-methyl-7-phenyl-N-p- tolylpyrazolo[1,5-a] pyrimidine-3-carboxamide205.7502 6.585 MLS000663265-01

N-(5-bromo-2- hydroxyphenyl)- 5-cyclopropyl-7-(difluoromethyl)pyrazolo[1,5- a]pyrimidine-3-carboxamide 2.36 Method 2MLS000716072-01

N-(3-hydroxyphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 1.86 Method 2 NCGC00182126- 01

N,N-dicyclohexyl-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.84 Method 3NCGC00182127- 01

N-(4-bromophenyl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.60 Method 3NCGC00182128- 01

5-cyclopropylidene-N-(2,4- dichlorophenyl)-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.78 Method 3NCGC00182129- 01

N-(4-acetylphenyl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.35 Method 3NCGC00182130- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(3,4- dimethoxyphenethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.25 Method 3NCGC00182131- 01

(5-cyclopropylidene-7- (difluoromethyl)-4,5- dihydropyrazolo[1,5-a]pyrimidin-3-yl)(3,4- dihydroisoquinolin-2(1H)- yl)methanone 2.37Method 3 NCGC00182134- 01

N-(4-chlorophenyl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.57 Method 3NCGC00182135- 01

5-cyclopropylidene-7- (difluoromethyl)- N-(pyridin-4-yl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 1.71 Method 3NCGC00182136- 01

5-cyclopropylidene-7- (difluoromethyl)- N-(pyridin-3-yl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 1.73 Method 3NCGC00182137- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(1- methylpiperidin-4-yl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 1.65 Method 3NCGC00182138- 01

5-cyclopropylidene-7- (difluoromethyl)- N-((1-methyl-1H-pyrazol-4-yl)methyl)-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 1.99 Method 3 NCGC00182140- 01

N-(4-fluorobenzyl)-5- methylene-7- (trifluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.29 Method 3NCGC00182143- 01

7-(difluoromethyl)-N- (4-methoxybenzyl)-5- methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.21 Method 3NCGC00182149- 01

7-(difluoromethyl)-N-(3,4- dimethoxyphenethyl)-5- methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.16 Method 3NCGC00182150- 01

N-benzyl-7- (difluoromethyl)-5- methylene-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 2.23 Method 3 NCGC00182153- 01

N-(4-bromo-2- (trifluoromethyl)phenyl)-7- (difluoromethyl)- 5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide NCGC00182155- 01

(7-(difluroomethyl)-5- methylene-4,5- dihydropyrazolo[1,5-a]pyrimidin-3-yl)(piperidin-1- yl)methanone 2.09 Method 3 NCGC00182156-01

7-(difluoromethyl)-N-(4- fluorobenzyl)-5- methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.24 Method 3NCGC00182157- 01

N-(4-chlorobenzyl)-7- (difluoromethyl)-5-methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.34 Method 3NCGC00182158- 01

7-(difluoromethyl)-N-(3,4- dimethoxybenzyl)-5- methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide NCGC00182163- 01

(7-(difluoromethyl)-5- methylene-4,5- dihydropyrazolo[1,5-a]pyrimidin-3-yl)(3,4- dihydroisoquinolin-2(1H)- yl)methanone 2.23Method 3 NCGC00182164- 01

N-allyl-7-methyl- 5-methylene- 4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 2.00 Method 3 NCGC00182168- 01

(5-cyclopropylidene-7- (difluoromethyl)-4,5- dihydropyrazolo[1,5-a]pyrimidin-3-yl)(indolin-1- yl)methanone 2.39 Method 3 NCGC00182169- 01

N-(5-bromopyridin-2-yl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.62 Method 3NCGC00182173- 01

N-(2-chloropyridin-3-yl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.43 Method 3NCGC00182177- 01

(5-cyclopropylidene-7- (difluoromethyl)-4,5- dihydropyrazolo[1,5-a]pyrimidin-3-yl)(2- methylpiperidin-1- yl)methanone 2.35 Method 3NCGC00182178- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(3,4- difluorophenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.51 Method 3NCGC00182180- 01

5-cyclopropylidene-7- (difluoromethyl)-N- (naphthalen-1-yl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.58 Method 3NCGC00182181- 01

5-cyclopropylidene-N-(3,4- difluorophenyl)-7- (trifluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.56 Method 3NCGC00182183- 01

5-cyclopropylidene-N-(3,5- dichlorophenyl)-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.77 Method 3NCGC00182184- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(4- nitrophenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.46 Method 3NCGC00182187- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(4- sulfamoylphenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.07 Method 3NCGC00182189- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(2- fluorophenyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.52 Method 3NCGC00182191- 01

N-(4-bromo-2- chlorophenyl)-5- cyclopropylidene-7- (difluoromethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide NCGC00182192- 01

(5-cyclopropylidene-7- (difluoromethyl)-4,5- dihydropyrazolo[,15-a]pyrimidin-3-yl) (6-methyl-3,4- dihydroquinolin-1(2H)- yl)methanone2.49 Method 3 NCGC00182193- 01

5-cyclopropylidene-7- (difluoromethyl)-N-(1- (pyridin-4-yl)ethyl)-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 1.71 Method 3NCGC00182194- 01

7-methyl-5-methylene- N-(3-(3- (trifluoromethyl)phenyl-carbamoyl)phenyl)-4,5- dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide2.5 Method 3 NCGC00182195- 01

N-benzyl-7-methyl-5- methylene-N-phenethyl-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 2.47 Method 3 NCGC00182196- 01

N-(4-acetamido-3- chlorophenyl)-7-methyl-5- methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide 2.07 Method 3NCGC00182199- 01

N-(2-hydroxyphenyl)-7- methyl-5-methylene-4,5- dihdyropyrazolo[1,5-a]pyrimidine-3-carboxamide 2.07 Method 3 NCGC00182200- 01

N-(isoquinolin-5-yl)-7- methyl-5-methylene-4,5- dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide 1.68 Method 3 NCGC00182241- 01

7-(difluoromethyl)-N-(2- fluorobenzyl)-5- methylene-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide NCGC00182242- 01

5-cyclopropylidene-7- (difluoromethyl)- N-p-tolyl-4,5-dihydropyrazolo[1,5- a]pyrimidine-3-carboxamide NCGC00187966- 01

N-(4-hydroxybutyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide3.717 NCGC00187968- 01

(5,7-dimethylpyrazolo[1,5- a]pyrimidin-3-yl)(4-(4-nitrophenyl)piperazin-1- yl)methanone 5.079 NCGC00187972- 01

N-isopropyl-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide 4.655NCGC00187973- 01

(5,7-dimethylpyrazolo[1,5- a]pyrimidin-3-yl)(pyrrolidin- 1-yl)methanone3.843 NCGC00187974- 01

N-(3-cyanophenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide5.472 NCGC00187977- 01

N,5,7-trimethylpyrazolo[1,5- a]pyrimidine-3-carboxamide 3.709NCGC00187979- 01

N-(2,2-difluoroethyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 4.386 NCGC00188757- 01

(5,7-dimethylpyrazolo[1,5- a]pyrimidine-3-yl)(4- methylpiperazin-1-yl)methanone 2.543 NCGC00188760- 01

5,7-dimethyl-N-(4- (methylsulfonyl) phenyl)pyrazolo [1,5-a]pyrimidine-3-carboxamide 4.759 NCGC00188761- 01

5,7-dimethyl-N-(pyridin-4- yl)pyrazolo[1,5-a] pyrimidine-3-carboxamide3.326 NCGC00188765- 01

N-(2-acetamidophenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 4.169 NCGC00188767- 01

N-(4-acetamidophenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 4.385 NCGC00188775- 01

5,7-dimethyl-N-(3,4,5- trimethoxyphenyl) pyrazolo[1,5-a]pyrimidine-3-carboxamide 5.239 NCGC00188777- 01

N-(3,4-dimethoxyphenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide 5.026 NCGC00229709- 01

N-(4-methoxyphenyl)-5- methyl-7-phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 6.165 NCGC00229710- 01

N-(3,4-dichlorophenyl)-5- methyl-7-phenylpyrazolo[1,5-a]pyrimidine-3-carboxamide 7.215 NCGC00229720- 01

5-methyl-7-phenyl-N-(2- (phenylthio)phenyl)pyrazolo [1,5-a]pyrimidine-3-carboxamide 7.281 CCB2-5-1

N-((2E,4E)-hexa-2,4-dien-2- yl)-5-methyl-7,8-dihydro-6H-cyclopenta[e]pyrazolo[1,5- a]pyrimidine-3-carboxamide CCB2-6-1

N-(4-ethylphenyl)-5-methyl- 7,8-dihydro-6H- cyclopenta[e]pyrazolo[1,5-a]pyrimidine-3-carboxamide CCB2-7-1

5-methyl-N-(4- (trifluoromethyl)phenyl)-7,8- dihydro-6H-cyclopenta[e]pyrazolo[1,5- a]pyrimidine-3-carboxamide CGB2-8-1

N-(3,4-dichlorophenyl)-5- methyl-7,8-dihydro-6H-cyclopenta[e]pyrazolo[1,5- a]pyrimidine-3-carboxamide CGB2-9-1

N-(4-ethynylphenyl)-5-methyl- 7.8-dihydro-6H- cyclopenta[e]pyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-10-1

N-(4-tert-butylcyclohexyl)-5- methyl-7,8-dihydro-6H-cyclopenta[e]pyrazolo[1,5- a]pyrimidine-3-carboxamide CGB2-12-1

5-methyl-N-p-tolyl-6,7,8,9- tetrahydropyrazolo[1,5-a]quinazoline-3-carboxamide CGB2-13-1

N-(4-ethylphenyl)-5-methyl- 6,7,8,9-tetrahydro- pyrazolo[1,5-a]quinazoline-3-carboxamide CGB2-14-1

5-methyl-N-(4- (trifluoromethyl)phenyl)- 6,7,8,9-tetrahydro-pyrazolo[1,5- a]quinazoline-3-carboxamide CGB2-16-1

N-(4-ethynylphenyl)- 5-methyl-6,7,8,9- tetrahydropyrazolo[1,5-a]quinazoline-3-carboxamide CGB2-17-1

N-(4-tert-butylcyclohexyl)-5- methyl-6,7,8,9- tetrahydropyrazolo[1,5-a]quinazoline-3-carboxamide CGB2-18-1

7-(difluoromethyl)- 5-methyl-N- p-tolylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-19-1

7-(difluoromethyl)-N-(4- ethylphenyl)-5- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-20-1

7-(difluoromethyl)- 5-methyl-N- (4-(trifluoromethyl)phenyl)pyrazolo[1,5-a]pyrimidine-3- carboxamide CGB2-21-1

N-(3,4-dichlorophenyl)-7- (difluoromethyl)-5- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-22-1

7-(difluoromethyl)-N-(4- ethynylphenyl)-5- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-23-1

N-(4-tert-butylcyclohexyl)-5- (difluoromethyl)-7- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-32-1

5,7-dimethyl-N-(4- morpholinophenyl) pyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-33-1

5,7-dimethyl-N- (4-(piperidin-1- yl)phenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-34-1

N-(3-(dimethyl- amino)phenyl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-35-1

N-(4-(dimethyl- amino)phenyl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-36-1

N-(4-bromophenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamideCGB2-39-1

N-(4′-cyanobiphenyl- 4-yl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-41-1

N-(3′-methoxybiphenyl-4-yl) 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-42-1

N-(3′-cyanobiphenyl- 4-yl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-43-1

N-(4′-(dimethyl- amino)biphenyl-4-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-62-1

N-(4′-methoxybiphenyl-4-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-63-1

N-(3′-(dimethyl- amino)biphenyl-4-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-68-1

N-(4-iodophenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamideCGB2-55-1

N-(3-bromophenyl)-5,7- dimethylpyrazolo[1,5- a]pyrimidine-3-carboxamideCGB2-56-1

N-(4′-methoxybiphenyl-3-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-57-1

N-(4′-cyanobiphenyl- 3-yl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-58-1

N-(3′-methoxybiphenyl- 3-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-59-1

N-(4′-(dimethyl- amino)biphenyl-3-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-60-1

N-(3′-(dimethyl- amino)biphenyl-3-yl)- 5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-61-1

5,7-dimethyl-N-(3-(pyrimidin- 5-yl)phenyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-67-1

N-(3′-cyanobiphenyl- 3-yl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-64-1

N-(4-ethylphenyl)- 5-methyl-7- (4-methylpiperazin-1- yl)pyrazolo[1,5-a]pyrimidine-3- carboxamide CGB2-65-1

N-(4-ethylphenyl)-5- methyl-7-(2- morpholinoethyl- amino)pyrazolo[1,5-a]pyrimidine-3- carboxamide CGB2-75-1

N-(4-ethylphenyl)- 5-methyl-7-(piperidin- 1-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-77-1

N-(4-ethylphenyl)- 5-methyl-7-morpholino- pyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-79-1

N-(4-ethylphenyl)-7-(4- isopropylpiperazin-1-yl)-5- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-80-1

N-(4-ethylphenyl)-7-(4- isobutylpiperazin-1-yl)-5- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-81-1

7-(4-(2-(dimethyl- amino)ethyl)piperazin- 1-yl)-N-(4-ethylphenyl)-5-methylpyrazolo[1,5- a]pyrimidine-3-carboxamide CGB2-82-1

7-(3,5-dimethyl- morpholino)-N- (4-ethylphenyl)-5- methylpyrazolo[1,5-a]pyrimidine-3-carboxamide CGB2-83-1

7-(3,5-dimethyl- piperazin-1-yl)- N-(4-ethylphenyl)-5-methylpyrazolo[1,5- a]pyrimidine-3-carboxamide CGB2-70-1

N-(4-(3-(dimethylamino) prop-1-ynyl)phenyl)-5,7- dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide

1-37. (canceled)
 38. A compound of the formula

or a pharmaceutically acceptable salt thereof, wherein R₁ ispyrimidinyl) C₀-C₄alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl,(pyrazolyl)C₀-C₂alkyl, (pyrrolyl)C₀-C₂alkyl, (imidazolyl)C₀-C₂alkyl,(thienyl)C₀-C₂alkyl, (oxazolyl)C₀-C₂alkyl, (thiazolyl)C₀-C₂alkyl,pyrolidinyl, naphthyl, quinolinyl, isoquinolinyl, tetrahydronaphthyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl tetrahydrofuranyl,morpholinyl, piperidinyl, or thiomorpholinyl, each of which isunsubstituted or substituted with one or more substituents independentlychosen from halogen, hydroxyl, cyano, nitro, amino, —CHO, —COOH,C₁-C₆alkyl, C₁-C₆alkoxy, C₂-C₆alkanoyl, mono- or di-C₁-C₆alkylamino,mono- or di-C₁-C₆alkylcarboxamide, C₁-C₆alkylester, C₁-C₆alkylthio,C₁-C₆alkylsulfonyl, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, and with 0 or 1substituents chosen from Y—Z— where Z is a covalent bond, C₁-C₄alkylene,—S—, —O—, —NR—, —C(O)—, —NHC(O)—, or —C(O)NH—, where R is hydrogen orC₁-C₄alkyl, and Y is phenyl or pyridyl, each of which is unsubstitutedor substituted with 1 to 3 substituents independently chosen fromhalogen, hydroxyl, cyano, nitro, amino, C₁-C₄alkyl, and C₁-C₄alkoxy; R₂is hydrogen or methyl; R₃ and R₆ are both hydrogen; and R₅ and R₇ areboth methyl; or one of R₅ and R₇ is methyl and the other phenyl; or oneof R₅ and R₇ is methyl and the other is difluoromethyl; with the provisothat R₁ is not unsubstituted cyclohexyl.
 39. The compound orpharmaceutically acceptable salt thereof of claim 38, wherein R₅ and R₇are both methyl.
 40. The compound or pharmaceutically acceptable saltthereof of claim 38, wherein the compound is selected from:N-(4-tert-butylcyclohexyl)-5,7-dimethylpyrazolo[1,5-a]pyrimidine-3-carboxamide;5,7-dimethyl-N-(quinolin-3-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide;5,7-dimethyl-N-(quinolin-2-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide;andN-(isoquinolin-5-yl)-7-methyl-5-methylene-4,5-dihydropyrazolo[1,5-a]pyrimidine-3-carboxamide.41. A pharmaceutical composition comprising the compound of claim 38together with a pharmaceutically acceptable carrier.
 42. Thepharmaceutical composition of claim 41, wherein the composition is inthe form of an oral dosage form.
 43. A method of treating Gaucherdisease in a patient or preventing the symptoms of Gaucher disease in apatient having a GBA gene mutation comprising administering atherapeutically effective amount of a compound or salt of claim 38 tothe patient.
 44. A method of increasing the amount of betaglucocerebrosidase in the white blood cells of patient having a GBA genemutation by administering an effective amount of a compound or salt ofclaim 38 to the patient.